The unique structure and biochemical composition of articular cartilage determine its biomechanical properties and in turn, its functionality. Current clinical techniques available to repair damaged cartilage are unable to reliably regenerate mechanically stable, functional tissue. Hence there is a need to identify new and effective means to repair damaged cartilage. The underpinning concept and long term aim of this research is that of "acellular repair", to produce a direct replacement for damaged cartilage using acellular cartilage-like constructs developed via tissue engineering principals: I) seeding of suitable cells onto scaffolds and culture in chondrogenic medium; 2) application of cyclic compressive loading to promote cell differentiation and deposition of cartilage-like matrix; 3) achievement of construct mechanical properties comparable to native cartilage and 4) construct decellularisation to avoid any immune response following implantation in to the patient. The aim of this thesis is to begin optimisation of the parameters required to deliver the acellular repair. Human foetal osteoblastic cells (1.19 cell line; hFOB), bovine synoviocytes and human bone marrow mesenchymal stem cells were investigated by seeding onto polyethylene terephthalate non-woven fibre scaffolds of differing porosities and cultured for 4 weeks in chondrogenic medium. Protein and DNA assays, plus scanning electron microscopy revealed that synoviocytes were the most effective in producing filled scaffolds with 90.2 % being the optimum scaffold porosity. Constructs were then subjected to 13 to 23 % cyclic compressive strain at 1 Hz for 1 hr per day, using an in-house bioreactor. After 56 days (sustained at Day 84) loaded constructs had compressive moduli comparable to the higher ranges of native cartilage and histological properties similar to cartilage itself. It is concluded that optimisation of the parameters to deliver the acellular repair concept has been achieved. Reliable production of mechanically functional constructs containing cartilage-like matrix will allow for the following stage, decellularisation, to be undertaken. The development of the acellular cartilage-like constructs, thus far, provides promise for potential future clinical application.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:612556 |
| Date | January 2012 |
| Creators | Finlay, Scott |
| Publisher | University of Leeds |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
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